InVivoPlus anti-mouse CD40L (CD154)

About InVivoPlus anti-mouse CD40L (CD154)

The MR-1 monoclonal antibody reacts with mouse CD154 also known as CD40 ligand. CD154 exists as a 39 kDa accessory molecule and belongs to the TNF superfamily of cytokines. CD154 is primarily expressed on the surface of activated CD4+ T lymphocytes but can also be expressed by platelets, mast cells, macrophages, basophils, NK cells, B lymphocytes, CD8+ T lymphocytes as well as non-hematopoietic cells including smooth muscle cells, endothelial cells, and epithelial cells. CD154 signals through CD40 and is thought to play a key role in T and B lymphocyte costimulation. The MR-1 monoclonal antibody has been reported to inhibit in vitro activation of B lymphocytes by blocking the binding of CD154 with CD40 on T helper cells as well as inhibit the formation of germinal centers and disrupt antigen-specific T cell responses. Additionally, the MR-1 antibody blocks interactions of T cells and antigen-presenting cells in vitro and blocks the development of experimental autoimmune disease in vivo.

The antibody solution should be stored undiluted at 4°C, and protected from prolonged exposure to light. Do not freeze.

Murine Pathogen Test Results

Mouse Norovirus: Negative

Mouse Parvovirus: Negative

Mouse Minute Virus: Negative

Mouse Hepatitis Virus: Negative

Reovirus Screen: Negative

Lymphocytic Choriomeningitis virus: Negative

Lactate Dehydrogenase-Elevating Virus: Negative

Mouse Rotavirus: Negative

Theiler's Murine Encephalomyelitis: Negative

Ectromelia/Mousepox Virus: Negative

Hantavirus: Negative

Polyoma Virus: Negative

Mouse Adenovirus: Negative

Sendai Virus: Negative

Mycoplasma Pulmonis: Negative

Pneumonia Virus of Mice: Negative

Mouse Cytomegalovirus: Negative

K Virus: Negative

RRID

AB_1107601

The InVivoPlus Difference

Our InVivoPlus™ antibodies feature all the great qualities of our InVivoMab™ antibodies. The InVivoPlus™ versions of our products are structurally and functionally identical to the InVivoMab™ versions with the added benefit of additional QC measures. InVivoPlus™ antibodies are screened for murine pathogens using ultrasensitive qPCR, screened for protein aggregation via dynamic light scattering, feature advanced binding validation via flow cytometry, ELISA, and/or Western blot, and are guaranteed to contain less than 1 endotoxin unit per milligram. Our InVivoPlus™ line of antibodies are designed to exceed the strict demands and rigorous standards required for in vivo work at any research organization.

Products related to InVivoPlus anti-mouse CD40L (CD154)

InVivoPlus anti-mouse CD154 (CD40L) (Clone: MR-1)

PU.1 is an ETS family transcription factor that is important for the development of multiple hematopoietic cell lineages. Previous work demonstrated a critical role for PU.1 in promoting Th9 development and in limiting Th2 cytokine production. Whether PU.1 has functions in other Th lineages is not clear. In this study, we examined the effects of ectopic expression of PU.1 in CD4+ T cells and observed decreased expression of genes involved with the function of T follicular helper (Tfh) cells, including Il21 and Tnfsf5 (encoding CD40L). T cells from conditional mutant mice that lack expression of PU.1 in T cells (Sfpi1lck-/-) demonstrated increased production of CD40L and IL-21 in vitro. Following adjuvant-dependent or adjuvant-independent immunization, we observed that Sfpi1lck-/- mice had increased numbers of Tfh cells, increased germinal center B cells (GCB cells), and increased Ab production in vivo. This correlated with increased expression of IL-21 and CD40L in Tfh cells from Sfpi1lck-/- mice compared with control mice. Finally, although blockade of IL-21 did not affect GCB cells in Sfpi1lck-/- mice, anti-CD40L treatment of immunized Sfpi1lck-/- mice decreased GCB cell numbers and Ag-specific Ig concentrations. Together, these data indicate an inhibitory role for PU.1 in the function of Tfh cells, germinal centers, and Tfh-dependent humoral immunity.

Brasseit, J., et al. (2015). "CD4 T cells are required for both development and maintenance of disease in a new mouse model of reversible colitis." Mucosal Immunol. doi: 10.1038/mi.2015.93. PubMed

Current therapies to treat inflammatory bowel diseases have limited efficacy, significant side effects, and often wane over time. Little is known about the cellular and molecular mechanisms operative in the process of mucosal healing from colitis. To study such events, we developed a new model of reversible colitis in which adoptive transfer of CD4+CD45RBhi T cells into Helicobacter typhlonius-colonized lymphopenic mice resulted in a rapid onset of colonic inflammation that was reversible through depletion of colitogenic T cells. Remission was associated with an improved clinical and histopathological score, reduced immune cell infiltration to the intestinal mucosa, altered intestinal gene expression profiles, regeneration of the colonic mucus layer, and the restoration of epithelial barrier integrity. Notably, colitogenic T cells were not only critical for induction of colitis but also for maintenance of disease. Depletion of colitogenic T cells resulted in a rapid drop in tumor necrosis factor alpha (TNFalpha) levels associated with reduced infiltration of inflammatory immune cells to sites of inflammation. Although neutralization of TNFalpha prevented the onset of colitis, anti-TNFalpha treatment of mice with established disease failed to resolve colonic inflammation. Collectively, this new model of reversible colitis provides an important research tool to study the dynamics of mucosal healing in chronic intestinal remitting-relapsing disorders.

Tissue effector cells of the monocyte lineage can differentiate into different cell types with specific cell function depending on their environment. The phenotype, developmental requirements, and functional mechanisms of immune protective macrophages that mediate the induction of transplantation tolerance remain elusive. Here, we demonstrate that costimulatory blockade favored accumulation of DC-SIGN-expressing macrophages that inhibited CD8(+) T cell immunity and promoted CD4(+)Foxp3(+) Treg cell expansion in numbers. Mechanistically, that simultaneous DC-SIGN engagement by fucosylated ligands and TLR4 signaling was required for production of immunoregulatory IL-10 associated with prolonged allograft survival. Deletion of DC-SIGN-expressing macrophages in vivo, interfering with their CSF1-dependent development, or preventing the DC-SIGN signaling pathway abrogated tolerance. Together, the results provide new insights into the tolerogenic effects of costimulatory blockade and identify DC-SIGN(+) suppressive macrophages as crucial mediators of immunological tolerance with the concomitant therapeutic implications in the clinic.

Transplantation is a cure for end-stage organ failure but, in the absence of pharmacological immunosuppression, allogeneic organs are acutely rejected. Such rejection invariably results in allosensitization and accelerated rejection of secondary donor-matched grafts. Transplantation tolerance can be induced in animals and a subset of humans, and enables long-term acceptance of allografts without maintenance immunosuppression. However, graft rejection can occur long after a state of transplantation tolerance has been acquired. When such an allograft is rejected, it has been assumed that the same rules of allosensitization apply as to non-tolerant hosts and that immunological tolerance is permanently lost. Using a mouse model of cardiac transplantation, we show that when Listeria monocytogenes infection precipitates acute rejection, thus abrogating transplantation tolerance, the donor-specific tolerant state re-emerges, allowing spontaneous acceptance of a donor-matched second transplant. These data demonstrate a setting in which the memory of allograft tolerance dominates over the memory of transplant rejection.

Memory CD8(+) T cells are programmed during the primary response for robust secondary responsiveness. Here we show that CD8(+) T cells responding to different epitopes of influenza virus received qualitatively different signals during the primary response that altered their secondary responsiveness. Nucleoprotein (NP)-specific CD8(+) T cells encountered antigen on CD40-licensed, CD70-expressing, CD103(-)CD11b(hi) dendritic cells (DCs) at later times in the primary response. As a consequence, they maintained CD25 expression and responded to interleukin-2 (IL-2) and CD27, which together programmed their robust secondary proliferative capacity and interferon-gamma (IFN-gamma)-producing ability. In contrast, polymerase (PA)-specific CD8(+) T cells did not encounter antigen-bearing, CD40-activated DCs at later times in the primary response, did not receive CD27 and CD25 signals, and were not programmed to become memory CD8(+) T cells with strong proliferative and cytokine-producing ability. As a result, CD8(+) T cells responding to abundant antigens, like NP, dominated the secondary response.

Effector and memory T cells may cross-react with allogeneic Ags to mediate graft rejection. Whereas the costimulation properties of Th1 cells are well studied, relatively little is known about the costimulation requirements of microbe-elicited Th17 cells. The costimulation blocker CTLA-4 Ig has been ineffective in the treatment of several Th17-driven autoimmune diseases and is associated with severe acute rejection following renal transplantation, leading us to investigate whether Th17 cells play a role in CD28/CTLA-4 blockade-resistant alloreactivity. We established an Ag-specific model in which Th1 and Th17 cells were elicited via Mycobacterium tuberculosis and Candida albicans immunization, respectively. C. albicans immunization elicited a higher frequency of Th17 cells and conferred resistance to costimulation blockade following transplantation. Compared with the M. tuberculosis group, C. albicans-elicited Th17 cells contained a higher frequency of IL-17(+)IFN-gamma(+) producers and a lower frequency of IL-10(+) and IL-10(+)IL-17(+) cells. Importantly, Th17 cells differentially regulated the CD28/CTLA-4 pathway, expressing similarly high CD28 but significantly greater amounts of CTLA-4 compared with Th1 cells. Ex vivo blockade experiments demonstrated that Th17 cells are more sensitive to CTLA-4 coinhibition and therefore less susceptible to CTLA-4 Ig. These novel insights into the differential regulation of CTLA-4 coinhibition on CD4(+) T cells have implications for the immunomodulation of pathologic T cell responses during transplantation and autoimmunity.

The developmental pathways of regulatory T cells (T(reg)) generation in the thymus are not fully understood. In this study, we reconstituted thymic development of Zap70-deficient thymocytes with a tetracycline-inducible Zap70 transgene to allow temporal dissection of T(reg) development. We find that T(reg) develop with distinctive kinetics, first appearing by day 4 among CD4 single-positive (SP) thymocytes. Accepted models of CD25(+)Foxp3(+) T(reg) selection suggest development via CD25(+)Foxp3(-) CD4 SP precursors. In contrast, our kinetic analysis revealed the presence of abundant CD25(-)Foxp3(+) cells that are highly efficient at maturing to CD25(+)Foxp3(+) cells in response to IL-2. CD25(-)Foxp3(+) cells more closely resembled mature T(reg) both with respect to kinetics of development and avidity for self-peptide MHC. These population also exhibited distinct requirements for cytokines during their development. CD25(-)Foxp3(+) cells were IL-15 dependent, whereas generation of CD25(+)Foxp3(+) specifically required IL-2. Finally, we found that IL-2 and IL-15 arose from distinct sources in vivo. IL-15 was of stromal origin, whereas IL-2 was of exclusively from hemopoetic cells that depended on intact CD4 lineage development but not either Ag-experienced or NKT cells.

T follicular helper (Tfh) cells provide help to B cells and are crucial for establishment of germinal center (GC) reactions, including production of high-affinity antibodies and generation of memory B cells and long-lived plasma cells. Here we report that the magnitude of the Tfh cell response was dictated by the amount of antigen and directly correlated with the magnitude of the GC B cell response. In addition, maintenance of the Tfh cell phenotype required sustained antigenic stimulation by GC B cells. In lymphopenic conditions, a strong and prolonged Tfh cell response led to bystander B cell activation, hypergammaglobulinemia, and production of poly- and self-reactive antibodies. These data demonstrate that antigen dose determines the size and duration of the Tfh cell response and GC reaction, highlight the transient nature of the Tfh cell phenotype, and suggest a link between overstimulation of Tfh cells and the development of dysregulated humoral immune responses.

We recently identified a protective MHC class Ib-restricted CD8 T cell response to infection with mouse polyomavirus. These CD8 T cells recognize a peptide from aa 139-147 of the VP2 viral capsid protein bound to the nonpolymorphic H-2Q9 molecule, a member of the Qa-2 family of beta(2)m-associated MHC class Ib molecules. Q9:VP2.139-specific CD8 T cells exhibit an unusual inflationary response characterized by a gradual expansion over 3 mo followed by a stable maintenance phase. We previously demonstrated that Q9:VP2.139-specific CD8 T cells are dependent on Ag for expansion, but not for long-term maintenance. In this study, we tested the hypothesis that the expansion and maintenance components of the Q9:VP2.139-specific T cell response are differentially dependent on CD4 T cell help and CD28 costimulation. Depletion of CD4(+) cells and CD28/CD40L blockade impaired expansion of Q9:VP2.139-specific CD8 T cells, and intrinsic CD28 signaling was sufficient for expansion. In contrast, CD4 T cell insufficiency, but not CD28/CD40L blockade, resulted in a decline in frequency of Q9:VP2.139-specific CD8 T cells during the maintenance phase. These results indicate that the Q9:VP2.139-specific CD8 T cell response to mouse polyomavirus infection depends on CD4 T cell help and CD28 costimulation for inflationary expansion, but only on CD4 T cell help for maintenance.

Memory B cells can be produced from the classical germinal center (GC) pathway or a less understood GC-independent route. We used antigen-based cell enrichment to assess the relative contributions of these pathways to the polyclonal memory B cell pool. We identified a CD38(+) GL7(+) B cell precursor population that differentiated directly into IgM(+) or isotype-switched (sw) Ig(+) memory B cells in a GC-independent fashion in response to strong CD40 stimulation. Alternatively, CD38(+) GL7(+) B cell precursors had the potential to become Bcl-6(+) GC cells that then generated primarily swIg(+) memory B cells. These results demonstrate that early IgM(+) and swIg(+) memory B cells are products of a GC-independent pathway, whereas later switched Ig(+) memory B cells are products of GC cells.

To design successful vaccines for chronic diseases, an understanding of memory CD8(+) T cell responses to persistent antigen restimulation is critical. However, most studies comparing memory and naive cell responses have been performed only in rapidly cleared acute infections. Herein, by comparing the responses of memory and naive CD8(+) T cells to acute and chronic lymphocytic choriomeningitis virus infection, we show that memory cells dominated over naive cells and were protective when present in sufficient numbers to quickly reduce infection. In contrast, when infection was not rapidly reduced, because of high antigen load or persistence, memory cells were quickly lost, unlike naive cells. This loss of memory cells was due to a block in sustaining cell proliferation, selective regulation by the inhibitory receptor 2B4, and increased reliance on CD4(+) T cell help. Thus, emphasizing the importance of designing vaccines that elicit effective CD4(+) T cell help and rapidly control infection.

Privacy Policy

Your privacy is important to us. To better protect your privacy, we provide this notice
explaining our online information practices and the choices you can make about the
way your information is collected and used. This notice applies to all information
collected or submitted on the Bio X Cell website.

We use the personal information collected from you in order to make ordering from our company more convenient. Such personal information as we collect is not used for any other purpose and is not shared.

We never use or share the personally identifiable information provided to us online in
ways unrelated to the ones described above.

To prevent unauthorized access, maintain data accuracy, and ensure the correct use
of information, we have put into place appropriate physical, electronic, and
managerial procedures to safeguard and secure the information we collect online.
You can correct factual errors in your personally identifiable information by sending
us a request detailing the error(s). To protect your privacy and security, we will also
take reasonable steps to verify your identity before granting access or making
corrections. In the unlikely event any of the personal information you supply us
online is misused, you may redress these issues by contacting us directly at customerservice@bxcell.com.
Should you have questions or concerns about this Privacy Policy, please call us at
1-603-298-8564 or send us an email at customerservice@bxcell.com.

Legal Disclaimer

All catalog products supplied by Bio X Cell are warranted, for a period no longer than twelve months, to perform to the specifications noted on our product information sheets. This warranty is conveyed to the original purchaser only and is not valid for products that have been modified in any way or handled in any way that is not in accordance to the noted storage and handling conditions. This further applies to any shipment that has undergone repackaging and/or reshipping.

It is the buyer’s responsibility to inspect the package upon arrival to ensure the correct product and size has been shipped, also to inspect the integrity of the product packaging for signs of damage from shipping. Bio X Cell will assume responsibility and immediately replace, upon product availability, any missing, incorrectly sized or damaged products so long as the report is made within two weeks of delivery receipt.

Any product that fails to perform as warranted, is eligible, at the discretion of Bio X Cell, to receive either a full replacement of the product or a full credit for the purchase price of the product. Any claim must be reported to us within twelve months from the date of receipt to qualify for a replacement or credit regardless of the date the product was used. We only warranty QC data as stated on our COA, not additional specs introduced by the client.

Bio X Cell reserves the right to discontinue the sale of any product at any time and may change product descriptions and handling instructions at any time without prior notice.

Bio X Cell does not extend any other warranty, whether expressed or implied, with respect to these products. The performance of the products is assumed completely by the buyer, and in no event, shall Bio X Cell be held liable for incidental, consequential or punitive damages.